Nanozymes are increasingly being used for agricultural applications, but their adoption is limited as they are generally considered toxic, have low cost-effectiveness, and pose complexity of fabrication. In this study, an organic compound-based, peroxidase-like nanozyme (OC nanozyme) was developed for use in the agricultural environment. This nanozyme was synthesized through a self-assembled one-pot particle synthesis process, interacting with urea and the metal ion to form a homogenous nanoparticle containing partially mimicked cofactors (Fe–N) of the natural enzyme. The OC nanozyme exhibited decent kinetic properties (H₂O₂/K_m:0.056 mM and V_max:2.19 μM s⁻¹) and pH stability. The OC nanozyme was successfully used to detect glyphosate via integrated colorimetric assay, with a good limit of detection (LOD) of at least 0.001 ng mL⁻¹. The authors envision that this agricultural-friendly OC nanozyme holds great potential for a wide range of agricultural applications.